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Title: Robust generation of human-chambered cardiac organoids from pluripotent stem cells for improved modelling of cardiovascular diseases
Authors: Ho, Beatrice Xuan
Pang, Jeremy Kah Sheng
Chen, Ying 
Loh, Yuin-Han 
An, Omer 
Yang, Henry He 
Seshachalam, Veerabrahma Pratap 
Koh, Judice LY
Chan, Woon-Khiong 
Ng, Shi Yan 
Soh, Boon Seng 
Keywords: Science & Technology
Life Sciences & Biomedicine
Cell & Tissue Engineering
Cell Biology
Medicine, Research & Experimental
Research & Experimental Medicine
Cardiac progenitor
Chambered cardiac organoid
Disease modelling
Human pluripotent stem cells
Single-cell RNA sequencing
Issue Date: 21-Dec-2022
Publisher: BMC
Citation: Ho, Beatrice Xuan, Pang, Jeremy Kah Sheng, Chen, Ying, Loh, Yuin-Han, An, Omer, Yang, Henry He, Seshachalam, Veerabrahma Pratap, Koh, Judice LY, Chan, Woon-Khiong, Ng, Shi Yan, Soh, Boon Seng (2022-12-21). Robust generation of human-chambered cardiac organoids from pluripotent stem cells for improved modelling of cardiovascular diseases. STEM CELL RESEARCH & THERAPY 13 (1). ScholarBank@NUS Repository.
Abstract: Background: Tissue organoids generated from human pluripotent stem cells are valuable tools for disease modelling and to understand developmental processes. While recent progress in human cardiac organoids revealed the ability of these stem cell-derived organoids to self-organize and intrinsically formed chamber-like structure containing a central cavity, it remained unclear the processes involved that enabled such chamber formation. Methods: Chambered cardiac organoids (CCOs) differentiated from human embryonic stem cells (H7) were generated by modulation of Wnt/ß-catenin signalling under fully defined conditions, and several growth factors essential for cardiac progenitor expansion. Transcriptomic profiling of day 8, day 14 and day 21 CCOs was performed by quantitative PCR and single-cell RNA sequencing. Endothelin-1 (EDN1) known to induce oxidative stress in cardiomyocytes was used to induce cardiac hypertrophy in CCOs in vitro. Functional characterization of cardiomyocyte contractile machinery was performed by immunofluorescence staining and analysis of brightfield and fluorescent video recordings. Quantitative PCR values between groups were compared using two-tailed Student’s t tests. Cardiac organoid parameters comparison between groups was performed using two-tailed Mann–Whitney U test when sample size is small; otherwise, Welch’s t test was used. Comparison of calcium kinetics parameters derived from the fluorescent data was performed using two-tailed Student’s t tests. Results: Importantly, we demonstrated that a threshold number of cardiac progenitor was essential to line the circumference of the inner cavity to ensure proper formation of a chamber within the organoid. Single-cell RNA sequencing revealed improved maturation over a time course, as evidenced from increased mRNA expression of cardiomyocyte maturation genes, ion channel genes and a metabolic shift from glycolysis to fatty acid ß-oxidation. Functionally, CCOs recapitulated clinical cardiac hypertrophy by exhibiting thickened chamber walls, reduced fractional shortening, and increased myofibrillar disarray upon treatment with EDN1. Furthermore, electrophysiological assessment of calcium transients displayed tachyarrhythmic phenotype observed as a consequence of rapid depolarization occurring prior to a complete repolarization. Conclusions: Our findings shed novel insights into the role of progenitors in CCO formation and pave the way for the robust generation of cardiac organoids, as a platform for future applications in disease modelling and drug screening in vitro.
ISSN: 1757-6512
DOI: 10.1186/s13287-022-03215-1
Appears in Collections:Staff Publications

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